Is there a dependence between expression of integrin receptors by peripheral blood immune cells and duration of tuberculous granuloma existence in the patients?

О. V. Berdyugina
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Abstract

Over recent years, the number of patients with tuberculosis has not decreased in the country and in worldwide. This is due to high resistance of the pathogen and changing mechanisms of bacterial perception by the human immune system thus requiring closer examination of the issue. Cell fusion during the formation of pulmonary tuberculous granuloma involves a large number of adhesive events. Importance of α1β1 integrin has been shown for the granuloma integrity during the chronic phase of infection. It has been proven that pulmonary tuberculous granuloma should be monitored, including with the detection of cells expressing CD11c, since they support the continuous priming of T cells at different stages of infection. The aim of this study was to answer the question, if there is a different expression of integrin receptors by immune cells from the patient’s peripheral blood at different stages of the existence of pulmonary tuberculous granuloma? The study involved 38 people: the first group (control) consisted of 15 practically healthy people; a second group included 11 subjects with pulmonary tuberculous granuloma; the condition was first diagnosed 2 to 10 months before the present study. A third group consisted of 12 patients with pulmonary tuberculous granuloma, with primary diagnosis established 12 to 219 months before this study. All the participants underwent a general clinical blood tests using a 5 Diff Mythic 22 AL analyzer (Cormay, Poland). The adhesion markers CD11b, CD11c were detected with a Coulter Epicx XL instrument (Beckman Coulter, USA). The following peripheral blood cell populations were determined: CD14- CD13lowCD11b+, CD14- CD13lowCD11c+, CD14+CD11b+, CD14+CD11c+, CD45+CD3- CD16+CD56+, CD45+CD3- CD16+CD56+CD11b+. Statistical processing of the results was performed in the Windows 10 operating environment (Microsoft Corp., USA), using Statistica v. 12.5 software (StatSoft, USA). Kruskal–Wallis one-way analysis of variance (pk-w), with differences significant at p < 0.017, as well as the Wald–Wolfowitz test (pw-w) at a significance level of p < 0.05 were used as criteria for assessing differences between the compared groups. In addition, cluster and factor analysis were implemented. When studying the role of β2-integrins, we have found that they play an important role in maintaining the existence of pulmonary tuberculous granuloma. An increase in total number of granulocytes, and CD11b-expressing granulocytes, a decrease in the population of lymphocytes, NK cells and NK cells expressing CD11c proved to be distinctive in cases of pulmonary tuberculous granuloma detected 0.5 years before the study. Characteristic changes observed in the study of peripheral blood in the patients with pulmonary tuberculous granuloma detected 9.5 years before the study were as follows: an increase in the leukocyte population, total monocyte number, as well as CD11band CD11c-expressing monocytes.
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患者外周血免疫细胞整合素受体的表达与结核性肉芽肿的存在时间之间是否存在依赖性?
近年来,该国和全世界的结核病患者人数没有减少。这是由于病原体的高耐药性和人类免疫系统细菌感知机制的变化,因此需要对该问题进行更深入的研究。肺结核性肉芽肿形成过程中的细胞融合涉及大量的粘附事件。在感染的慢性期,α1β1整合素对肉芽肿的完整性具有重要意义。已经证明,应该监测肺结核性肉芽肿,包括检测表达CD11c的细胞,因为它们支持T细胞在不同感染阶段的持续启动。本研究的目的是回答这样一个问题:在肺结核性肉芽肿存在的不同阶段,患者外周血的免疫细胞是否存在整合素受体的不同表达?这项研究涉及38人:第一组(对照组)由15名实际健康的人组成;第二组包括11例肺结核性肉芽肿患者;在本研究前2至10个月首次诊断出这种情况。第三组由12名肺结核性肉芽肿患者组成,在本研究前12至219个月确定了初步诊断。所有参与者都使用5 Diff Mythic 22 AL分析仪(波兰科梅)进行了一般临床血液测试。用Coulter Epicx XL仪器(Beckman Coulter,USA)检测粘附标记CD11b、CD11c。测定以下外周血细胞群:CD14-CD13低CD11b+、CD14-CD13低CD11c+、CD14+CD11b+、CD14+CD11c+,CD45+CD3-CD16+CD56+、CD45+CD3-CD16+CD56+CD11b+。结果的统计处理是在Windows 10操作环境(Microsoft Corp.,USA)中使用Statistica v.12.5软件(StatSoft,USA)进行的。Kruskal–Wallis单向方差分析(pk-w)和Wald–Wolfowitz检验(pw-w)被用作评估比较组之间差异的标准,前者的差异在p<0.017时具有显著性,后者的显著性水平为p<0.05。此外,还进行了聚类分析和因子分析。在研究β2-整合素的作用时,我们发现它们在维持肺结核性肉芽肿的存在中起着重要作用。在研究前0.5年检测到的肺结核性肉芽肿病例中,粒细胞总数和表达CD11b的粒细胞的增加,淋巴细胞、NK细胞和表达CD11c的NK细胞数量的减少被证明是显著的。在研究前9.5年检测到的肺结核性肉芽肿患者的外周血研究中观察到的特征变化如下:白细胞数量、单核细胞总数以及表达CD11b和CD11c的单核细胞增加。
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